Characterization of the physical and mechanical properties of femoral bone defects filled with polyanionic collagen scaffolds in ovariectomized rats

The aim of this study was to evaluate the effect of scaffolds native or polyanionic collagen matrix (submitted to alkaline treatment for 48 or 96 hours, PCM48 or PCM96, respectively) on the repair of osteoporosis bone fractures resulting from the gonadal hormone alterations caused by ovariectomy in rats undergoing hormone replacement therapy. The physical and mechanical characteristics of bone were analyzed. Macroscopic analysis revealed the absence of pathological alterations in the implanted areas. The percent mineral matter and bone mineral density of the femurs were lower in ovariectomized rats. The mechanical strength of newly formed bone was greater in the area receiving the PCM96 scaffolds compared to the area implanted with the native scaffolds. The PCM96 scaffold is the best choice for bone repair in animals with hormone deficiency since it promotes faster bone growth and good mechanical strength.23924

Bone Plasticity in Response to Exercise Is Sex-Dependent in Rats

Purpose: To characterize the potential sexual dimorphism of bone in response to exercise.Methods: Young male and female Wistar rats were either submitted to 12 weeks of exercise or remained sedentary. the training load was adjusted at the mid-trial (week 6) by the maximal speed test. A mechanical test was performed to measure the maximal force, resilience, stiffness, and fracture load. the bone structure, formation, and resorption were obtained by histomorphometric analyses. Type I collagen (COL I) mRNA expression and tartrate-resistant acid phosphatase (TRAP) mRNA expression were evaluated by quantitative real-time PCR (qPCR).Results: the male and female trained rats significantly improved their maximum speed during the maximal exercise test (main effect of training; p<0.0001). the male rats were significantly heavier than the females, irrespective of training (main effect of sex; p<0.0001). Similarly, both the weight and length of the femur were greater for the male rats when compared with the females (main effect of sex; p<0.0001 and p<0.0001, respectively). the trabecular volume was positively affected by exercise in male and female rats (main effect of training; p = 0.001), whereas the trabecular thickness, resilience, mineral apposition rate, and bone formation rate increased only in the trained males (within-sex comparison; p<0.05 for all parameters), demonstrating the sexual dimorphism in response to exercise. Accordingly, the number of osteocytes increased significantly only in the trained males (within-sex comparison; p<0.05). Pearson's correlation analyses revealed that the COL I mRNA expression and TRAP mRNA expression were positively and negatively, respectively, related to the parameters of bone remodeling obtained from the histomorphometric analysis (r = 0.59 to 0.85; p<0.05).Conclusion: Exercise yielded differential adaptations with respect to bone structure, biomechanical proprieties, and molecular signaling in male and female rats.Univ São Paulo, Sch Med, Div Nephrol, São Paulo, BrazilUniv São Paulo, Sch Phys Educ & Sport, Dept Sports, São Paulo, BrazilUniv São Paulo, Inst Biomed Sci, Dept Anat, São Paulo, BrazilUniv São Paulo, Sch Med, Div Rheumatol, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Physiol Sci, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Physiol Sci, São Paulo, BrazilWeb of Scienc

Abnormal thyroid hormone status differentially affects bone mass accrual and bone strength in C3H/HeJ mice: a mouse model of type I deiodinase deficiency

C3H/HeJ (C3H) mice are deficient of type I deiodinase (D1), an enzyme that activates thyroid hormone (TH), converting thyroxine (T4) to triiodothyronine (T3). Nevertheless, C3H mice present normal serum T3 and a gross euthyroid phenotype. To investigate if a global D1 deficiency interferes in the TH effects on bone, we compared bone growth, bone mass accrual and bone strength of C3H and C57BL/6J (B6) mice under abnormal TH status. Four-week-old female mice of both strains were grouped as Euthyroid, Hypothyroid (pharmacologically-induced), 1xT4 and 10xT4 (hypothyroid animals receiving 1- or 10-fold the physiological dose of T4 /day/16 weeks). Hypothyroidism and TH excess similarly impaired body weight (BW) gain and body growth in both mice strains. In contrast, whereas hypothyroidism only slightly impaired bone mineral density (BMD) accrual in B6 mice, it severely impaired BMD accrual in C3H mice. No differences were observed in serum and bone concentrations of T3 between hypothyroid animals of both strains. Interestingly, treatment with 10xT4 was less deleterious to BMD accrual in C3H than in B6 mice and resulted in less elevated T3 serum levels in B6 than in C3H mice, which is probably explained by the lower D1 activity in C3H mice. In addition, hypothyroidism decreased bone strength only in C3H but not in B6 mice, while TH excess decreased this parameter in both strains. These findings indicate that D1 deficiency contributes to the TH excess-induced differences in bone mass accrual in C3H vs. B6 mice and suggest that deiodinase-unrelated genetic factors might account for the different skeleton responses to hypothyroidism between strains

Proteogenomics connects somatic mutations to signalling in breast cancer

Somatic mutations have been extensively characterized in breast cancer, but the effects of these genetic alterations on the proteomic landscape remain poorly understood. We describe quantitative mass spectrometry-based proteomic and phosphoproteomic analyses of 105 genomically annotated breast cancers of which 77 provided high-quality data. Integrated analyses allowed insights into the somatic cancer genome including the consequences of chromosomal loss, such as the 5q deletion characteristic of basal-like breast cancer. The 5q trans effects were interrogated against the Library of Integrated Network-based Cellular Signatures, thereby connecting CETN3 and SKP1 loss to elevated expression of EGFR, and SKP1 loss also to increased SRC. Global proteomic data confirmed a stromal-enriched group in addition to basal and luminal clusters and pathway analysis of the phosphoproteome identified a G Protein-coupled receptor cluster that was not readily identified at the mRNA level. Besides ERBB2, other amplicon-associated, highly phosphorylated kinases were identified, including CDK12, PAK1, PTK2, RIPK2 and TLK2. We demonstrate that proteogenomic analysis of breast cancer elucidates functional consequences of somatic mutations, narrows candidate nominations for driver genes within large deletions and amplified regions, and identifies therapeutic targets

Detectable clonal mosaicism and its relationship to aging and cancer

In an analysis of 31,717 cancer cases and 26,136 cancer-free controls from 13 genome-wide association studies, we observed large chromosomal abnormalities in a subset of clones in DNA obtained from blood or buccal samples. We observed mosaic abnormalities, either aneuploidy or copy-neutral loss of heterozygosity, of >2 Mb in size in autosomes of 517 individuals (0.89%), with abnormal cell proportions of between 7% and 95%. In cancer-free individuals, frequency increased with age, from 0.23% under 50 years to 1.91% between 75 and 79 years (P = 4.8 × 10(-8)). Mosaic abnormalities were more frequent in individuals with solid tumors (0.97% versus 0.74% in cancer-free individuals; odds ratio (OR) = 1.25; P = 0.016), with stronger association with cases who had DNA collected before diagnosis or treatment (OR = 1.45; P = 0.0005). Detectable mosaicism was also more common in individuals for whom DNA was collected at least 1 year before diagnosis with leukemia compared to cancer-free individuals (OR = 35.4; P = 3.8 × 10(-11)). These findings underscore the time-dependent nature of somatic events in the etiology of cancer and potentially other late-onset diseases

Efeito da simpatectomia química sobre as vértebras de ratas osteopênicas induzidas pela ovariectomia

Osteoporosis is a pathology that afflicts mainly postmenopausal women, consequently this disorder generates severe social and economic consequences. This disease is characterized by low bone mass (osteopenia) and bone tissue microarchitectural deterioration, which leads to enhanced bone fragility, thus resulting in fracture risk increase. As an experimental mode of bone loss in post-menopausal women the ovariectomized rat has been used. The periosteum is innervated by sympathetic nervous system (SNS) whose neurotransmitters regulate direct and indirectly the bone metabolism. In this study, we evaluated the possible influence of SNS on physical, biometric, biomechanical and biochemical parameters of the 5th lumbar vertebrae of ovariectomized (ovx) rats. Therefore, 4-month-old female rats were randomized in 5 groups: Basal (sacrificed at the beginning of the experimental), Intact (treated with 0,9% saline, sc), Control (ovx rats treated with 0,9% saline, sc), GUA 30 (ovx rats treated with 37 mg of guanetidine /kg, sc, for 30 days) and GUA 8 (ovx rats treated with 40 mg of guanetidine /kg, sc, for 8 weeks). The animals were sacrificed at the end of 8 weeks, except for the Basal group. The 5th lumbar vertebra was removed and defleshed and physical, miometric, biomechanical and chemical properties were analyzed. The denervation was confirmed by palpebral ptose and diarrhea observed in all treated animals. The Control group showed lower mineral and bone densities than the Intact group with consequent lower biomechanical parameter, indicating that castration was effective to induce osteopenia. The sympathetic denervation partially inhibited the osteopenia as shown by the higher values of mineral and bone densities when compared with the Control group. But, these better physical parameters did not induced better bone quality as shown by biomechanics.Financiadora de Estudos e ProjetosA osteoporose é uma osteopatologia que atinge principalmente a mulheres após a menopausa, com graves conseqüências sociais e econômicas. Ela é caracterizada pela perda de massa óssea (osteopenia) seguida de alteração na microstrutura do tecido ósseo fazendo com que o indivíduo fique mais propenso a ter fraturas. A rata castrada é um modelo experimental de osteoporose bem aceito na literatura. O Sistema Nervoso Simpático (SNS) inerva o periósteo e seus neurotransmissors regulam diretamente e indiretamente o metabolismo ósseo. O objetivo deste trabalho foi verificar a influência do Sistema Nervoso Simpático sobre os parâmetros biométricos, biomecânico, físicos e bioquímicos da quinta vértebra lombar (L5) de ratas ovariectomizadas (OVX). Para isso foram uilizadas ratas de 4 meses de idade divididas em 5 grupos: Basal (sacrificado no início do experimento), Intacto (Salina 0,9% s.c.), Controle (OVX+Salina 0,9% s.c.), GUA 30 (OVX+guanetidina 37mg/kg.dia s.c. durante 30 dias) e GUA 8 (OVX+guanetidina 40mg/kg.dia s.c. durante 8 semanas). Os animais forma sacrificados ao final de 8 semanas, exceto o grupo basal, e suas vértebras L5 foram dissecadas de suas partes moles e submetidas a análises biométricas, biomecânica, físicas e químicas. O efeito da desnervação pela guanetidina foi observada através da ptose palpebral e diarréia em todos os animais tratados com guanetidina. O grupo Controle apresentou menores valores de densidade mineral e óssea do que o grupo intacto, ,que refletiu sobre a resposta biomecânica, indicando que a indução da osteopenia pela ovariectomia foi efetiva. A simpatectomia inibiu parcialmente o desenvolvimento da osteopenia, pois os parâmetros densidade mineral e ósseo foram maiores em relação ao grupo Controle, porém estes parâmetros não refletiram sobre a propriedade biomecânica

Efeitos da administração crônica de nanopartículas de óxido de ferro magnético em ratos adultos jovens

The fast development of the nanotechnology, and the use of this technology to develop new products to help in diagnosis diseases, drugs and treatments for many diseases is increasing. Some authors have been warning about the nanoparticles use, because the real affect, toxic or not, still unknown. Iron oxide nanoparticles have been used in contrast liquid for magnetic resonance imaging, cancer treatment, among others. The aim of this work was to analyze the possible effects of chronic treatment with these nanoparticles in adults rats. The animals were distributed in 3 groups: Control (saline 0.9% - 0.1 mL/100 of body weight), nFe 0.3 (magnetic iron oxide nanoparticles 0.3 mg/kg BW) and nFe 0.6 (magnetic iron oxide nanoparticles 0.6 mg/kg BW). The animals were treated by gastric gavage during 8 weeks, 5 days per week. The body weight of the animals from the nFe 0.6 group decreased when compared to animals from the control group, and the BW of the animals from nFe 0.3 group was not different from the control group. The animals from the nFe 0.6 group showed the higher blood levels of cholesterol, although the levels of glucose, urea, creatinine, AST, ALT and alkaline phosphatase, plus hematology were not different from the animals of control group. Moreover, the seminal gland and ventral prostate of the animals of nFe 0.6 were atrophied. All those parameters of animals from nFe 0.3 group were not different compared to control group. The bone (femur and vertebra) biometric, biomechanical and biophysical parameters were similar in the animals from the different experimental groups. These results showed that high concentrations of magnetic iron oxide nanoparticles could be toxic and dangerous to the rats when take daily, but in low concentration it could be safe.Financiadora de Estudos e ProjetosCom o rápido desenvolvimento da nanotecnologia, o homem vem utilizando cada vez mais essa nova tecnologia para auxiliá-lo no desenvolvimento de novos produtos que auxiliem no diagnóstico de doenças, fármacos e tratamentos de inúmeras enfermidades. Alguns autores vêm chamando a atenção para o uso de nanopartículas em vários produtos comerciais, pois pouco se sabe sobre sua toxicidade. Especificamente as nanopartículas de óxido de ferro magnético vêm sendo utilizadas em líquidos de contraste de ressonância magnética, tratamento de câncer, entre outros. Com isso, o objetivo deste trabalho foi analisar os possíveis efeitos tóxicos dessas nanopartículas administradas cronicamente em ratos adultos jovens. Ratos Wistar machos foram distribuídos em 3 grupos experimentais: Controle (salina 0,9% - 0,1mL/100g MC), nFe 0,3 (nanopartículas de óxido de ferro - 0,3 mg/kg MC) e nFe 0,6 (nanopartículas de óxido de ferro - 0,6 mg/kg MC). O tratamento foi realizado por gavagem gástrica, 5 dias por semana, durante 8 semanas. As análises de massa corporal (MC) dos animais indicou que a concentração de 0,6 mg/kg MC de nanopartículas de óxido de ferro magnético levou à uma diminuição da MC dos animais em relação aos animais tratados com 0,3 mg/kg MC e aos animais tratados solução salina 0,9%. Os animais do grupo nFe 0,6 apresentaram aumento de colesterol sanguíneo, entretanto as concentrações de glicose, ferro, uréia, creatinina, AST, ALT e fosfatase alcalina, além da análise hematológica não foram diferentes dos animais do grupo controle. Além disso, as glândulas seminais e próstatas ventrais dos animais tratados com nFe 0,6 mg/kg MC atrofiaram, sendo que as dos animais do grupo nFe 0,3 não apresentaram diferença em relação as do grupo controle. Os parâmetros biométricos, biomecânicos e biofísicos dos ossos (fêmur e vértebra) foram semelhantes nos 3 grupos experimentais. Portanto, o tratamento crônico com nanopartículas na concentração de 0,6 mg/kg MC foi prejudicial à saúde dos animais, exceto no tecido ósseo, sendo que tal efeito não foi observado com a concentração mais baixa

Efeito de um novo produto fermentado de soja, enriquecido com isoflavonas e cálcio, sobre o tecido ósseo de ratas

O objetivo deste estudo foi avaliar o efeito da ingestão de um produto a base de soja, fermentado com Enterococcus faecium e Lactobacillus jugurti, enriquecido com isoflavonas e cálcio sobre o peso corpóreo e sobre o tecido ósseo de ratas maduras ovariectomizadas. Os animais foram divididos em 5 grupos: pseudo-ovariectomizados, ovariectomizados, ovariectomizados que ingeriram produto fermentado de soja enriquecido com isoflavonas e cálcio, ovariectomizados que ingeriram produto fermentado de soja enriquecido com cálcio e ovariectomizados que ingeriram produto não fermentado enriquecido com cálcio. Tíbias e fêmures foram utilizados para análise do comprimento ósseo; ensaio mecânico de flexão de três pontos; densidade óssea (princípio de Arquimedes); conteúdo mineral ósseo; conteúdo de cálcio ósseo e medida das larguras das trabéculas. A ingestão do produto fermentado de soja enriquecido com isoflavonas e cálcio apesar de não ter exercido efeitos nos parâmetros como comprimento, densidade, conteúdo mineral, conteúdo de cálcio e carga máxima suportada, foi capaz de evitar o aumento de peso corpóreo causado pela ovariectomia, além de ter demonstrado uma tendência à proteção das trabéculas, prevenindo a redução da largura das mesmas ocasionada pela castração.Summary The objective was to evaluate the effect of soy fermented product intake on the corporal weight and bone tissue of ovariectomized mature rats. This product was fermented with Enterococcus faecium and Lactobacillus jugurti and enriched with isoflavones and calcium. The animals were divided in 5 groups: sham-ovariectomized; ovariectomized; ovariectomized treated with soy fermented product enriched with isoflavones and calcium; ovariectomized treated with soy fermented product enriched with calcium and ovariectomized treated with non-fermented product enriched only with calcium. In order to evaluate the effect of the tested product on bone tissue (femur and tibia), the following parameters were analyzed: length; mechanical assay of three points; density (Archimedes principle); mineral content; calcium content; measure of the trabecular widths. The corporal weight of group treated with soy fermented product containing isoflavones and calcium showed no statistical difference from sham-ovariectomized group and trabecular widths tended to have larger than ovariectomized group. However, there was no significant difference to the other evaluated parameters in result of the diverse treatments. Thus, soy fermented product enriched with isoflavones and calcium inhibited the increasing of corporal weight caused by ovariectomy and revealed a tendency to trabecular protection after castration

Efeito de um novo produto fermentado de soja, enriquecido com isoflavonas e cálcio, sobre o tecido ósseo de ratas

O objetivo deste estudo foi avaliar o efeito da ingestão de um produto a base de soja, fermentado com Enterococcus faecium e Lactobacillus jugurti, enriquecido com isoflavonas e cálcio sobre o peso corpóreo e sobre o tecido ósseo de ratas maduras ovariectomizadas. Os animais foram divididos em 5 grupos: pseudo-ovariectomizados, ovariectomizados, ovariectomizados que ingeriram produto fermentado de soja enriquecido com isoflavonas e cálcio, ovariectomizados que ingeriram produto fermentado de soja enriquecido com cálcio e ovariectomizados que ingeriram produto não fermentado enriquecido com cálcio. Tíbias e fêmures foram utilizados para análise do comprimento ósseo; ensaio mecânico de flexão de três pontos; densidade óssea (princípio de Arquimedes); conteúdo mineral ósseo; conteúdo de cálcio ósseo e medida das larguras das trabéculas. A ingestão do produto fermentado de soja enriquecido com isoflavonas e cálcio apesar de não ter exercido efeitos nos parâmetros como comprimento, densidade, conteúdo mineral, conteúdo de cálcio e carga máxima suportada, foi capaz de evitar o aumento de peso corpóreo causado pela ovariectomia, além de ter demonstrado uma tendência à proteção das trabéculas, prevenindo a redução da largura das mesmas ocasionada pela castração